Biotechnology LettersPub Date : 2024-10-01Epub Date: 2024-05-21DOI: 10.1007/s10529-024-03477-0
Mohaddeseh Salehghamari, Mansour Mashreghi, Maryam M Matin, Zeinab Neshati
{"title":"Development of a bacterial cellulose-gelatin composite as a suitable scaffold for cardiac tissue engineering.","authors":"Mohaddeseh Salehghamari, Mansour Mashreghi, Maryam M Matin, Zeinab Neshati","doi":"10.1007/s10529-024-03477-0","DOIUrl":"10.1007/s10529-024-03477-0","url":null,"abstract":"<p><strong>Purpose: </strong>Cardiac tissue engineering is suggested as a promising approach to overcome problems associated with impaired myocardium. This is the first study to investigate the use of BC and gelatin for cardiomyocyte adhesion and growth.</p><p><strong>Methods: </strong>Bacterial cellulose (BC) membranes were produced by Komagataeibacter xylinus and coated or mixed with gelatin to make gelatin-coated BC (BCG) or gelatin-mixed BC (mBCG) scaffolds, respectively. BC based-scaffolds were characterized via SEM, FTIR, XRD, and AFM. Neonatal rat-ventricular cardiomyocytes (nr-vCMCs) were cultured on the scaffolds to check the capability of the composites for cardiomyocyte attachment, growth and expansion.</p><p><strong>Results: </strong>The average nanofibrils diameter in all scaffolds was suitable (~ 30-65 nm) for nr-vCMCs culture. Pore diameter (≥ 10 µm), surface roughness (~ 182 nm), elastic modulus (0.075 ± 0.015 MPa) in mBCG were in accordance with cardiomyocyte requirements, so that mBCG could better support attachment of nr-vCMCs with high concentration of gelatin, and appropriate surface roughness. Also, it could better support growth and expansion of nr-vCMCs due to submicron scale of nanofibrils and proper elasticity (~ 0.075 MPa). The viability of nr-vCMCs on BC and BCG scaffolds was very low even at day 2 of culture (~ ≤ 40%), but, mBCG could promote a metabolic active state of nr-vCMCs until day 7 (~ ≥ 50%).</p><p><strong>Conclusion: </strong>According to our results, mBCG scaffold was the most suitable composite for cardiomyocyte culture, regarding its physicochemical and cell characteristics. It is suggested that improvement in mBCG stability and cell attachment features may provide a convenient scaffold for cardiac tissue engineering.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"887-905"},"PeriodicalIF":2.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141070298","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Biotechnology LettersPub Date : 2024-10-01Epub Date: 2024-06-20DOI: 10.1007/s10529-024-03506-y
Luiza Carolina França Opretzka, Cláudio Damasceno Pinto, Jânio Rodrigo de Jesus Santos, Alyne Almeida de Lima, Milena Botelho Pereira Soares, Cristiane Flora Villarreal
{"title":"Mesenchymal stem cell-derived cell-free technologies: a patent landscape.","authors":"Luiza Carolina França Opretzka, Cláudio Damasceno Pinto, Jânio Rodrigo de Jesus Santos, Alyne Almeida de Lima, Milena Botelho Pereira Soares, Cristiane Flora Villarreal","doi":"10.1007/s10529-024-03506-y","DOIUrl":"10.1007/s10529-024-03506-y","url":null,"abstract":"<p><p>Mesenchymal stem/stromal cells (MSC) play a pivotal role in regenerative therapies. Recent studies show that factors secreted by MSC can replicate their biological activity, driving the emergence of cell-free therapy, likely to surpass stem cell therapy. Patents are an objective measure of R&D and innovation activities, and patent mapping allows us to verify the state of the art and technology, anticipate trends, and identify emerging lines of research. This review performed a search on Derwent World Patents Index™ and retrieved 269 patent families related to the MSC-derived cell-free products. Analysis reveals an exponential increase in patents from the mid-2010s, primarily focusing on exosomes. The patent's contents offer a great diversity of applications and associated technologies by using the products as medicinal agents or drug delivery systems. Nevertheless, numerous application branches remain unexplored, suggesting vast potential for cell-free technologies alone or combined with other approaches.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"907-924"},"PeriodicalIF":2.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141426259","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Biotechnology LettersPub Date : 2024-10-01Epub Date: 2024-05-08DOI: 10.1007/s10529-024-03489-w
Marcus Geissler, Christoph Neubauer, Yuriy V Sheludko, Adrian Brückner, Heribert Warzecha
{"title":"Nepeta cataria L. (catnip) can serve as a chassis for the engineering of secondary metabolic pathways.","authors":"Marcus Geissler, Christoph Neubauer, Yuriy V Sheludko, Adrian Brückner, Heribert Warzecha","doi":"10.1007/s10529-024-03489-w","DOIUrl":"10.1007/s10529-024-03489-w","url":null,"abstract":"<p><strong>Objective: </strong>Evaluation of Nepeta cataria as a host with specific endogenous metabolite background for transient expression and metabolic engineering of secondary biosynthetic sequences.</p><p><strong>Results: </strong>The reporter gene gfp::licBM3 as well as three biosynthetic genes leading to the formation of the cannabinoid precursor olivetolic acid were adopted to the modular cloning standard GoldenBraid, transiently expressed in two chemotypes of N. cataria and compared to Nicotiana benthamiana. To estimate the expression efficiency in both hosts, quantification of the reporter activity was carried out with a sensitive and specific lichenase assay. While N. benthamiana exhibited lichenase activity of 676 ± 94 μmol g<sup>-1</sup> s<sup>-1</sup>, N. cataria cultivar '1000', and the cultivar 'Citriodora' showed an activity of 37 ± 8 μmol g<sup>-1</sup> s<sup>-1</sup> and 18 ± 4 μmol g<sup>-1</sup> s<sup>-1</sup>, respectively. Further, combinatorial expression of genes involved in cannabinoid biosynthetic pathway acyl-activating enzyme 1 (aae1), olivetol synthase (ols) and olivetolic acid cyclase (oac) in N. cataria cv. resulted presumably in the in vivo production of olivetolic acid glycosides.</p><p><strong>Conclusion: </strong>Nepeta cataria is amenable to Agrobacterium-mediated transient expression and could serve as a novel chassis for the engineering of secondary metabolic pathways and transient evaluation of heterologous genes.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"843-850"},"PeriodicalIF":2.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11415451/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140875752","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Biotechnology LettersPub Date : 2024-10-01Epub Date: 2024-06-25DOI: 10.1007/s10529-024-03508-w
Shumin Hu, Qingqing Qin, Cui Zhang, Junhong Yu, Shuli Huang, Jia Liu, Zhaoxia Yang
{"title":"The effect of L-cysteine on starch and protein degradation during barley germination.","authors":"Shumin Hu, Qingqing Qin, Cui Zhang, Junhong Yu, Shuli Huang, Jia Liu, Zhaoxia Yang","doi":"10.1007/s10529-024-03508-w","DOIUrl":"10.1007/s10529-024-03508-w","url":null,"abstract":"<p><strong>Objectives: </strong>In order to investigate the impact of L-cysteine (L-Cys) on starch and protein degradation during barley germination. The amylase activities, degradation of macromolecules during germination were determined in this study.</p><p><strong>Methods: </strong>Barley was germinated in petri dish for 0 to 5 days with different levels of L-Cys (0 mM, 2.5 mM, 5 mM, 10 mM).</p><p><strong>Results: </strong>L-Cys addition increased the total limit dextrinase (LD) activities and decreased the LD inhibitor activities during whole germination stage. The activities of α-amylase, β-amylase and free LD were increased with the addition of 2.5, 5 mM L-Cys at germination days 1 to 4. Due to higher amylase in malt with the addition of L-Cys, the non-fermentable sugars were reduced and the glucose, maltotriose were improved. Furthermore, the protein degradation analysis showed that low molecular weight protein increased and middle molecular weight protein decreased obviously in wort from the malt germinated with L-Cys, demonstrating that the L-Cys promote the protein degradation. Lastly, the filtration performance of malt with the addition of L-Cys during malting was better than the control.</p><p><strong>Conclusion: </strong>In conclusion, L-Cys can promote the degradation of storage material (starch, protein) during barley germination, leading to a better green malt quality.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"861-870"},"PeriodicalIF":2.0,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141445355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Nitrile hydratase as a promising biocatalyst: recent advances and future prospects","authors":"Chao Feng, Jing Chen, Wenxin Ye, Zhanshi Wang","doi":"10.1007/s10529-024-03530-y","DOIUrl":"https://doi.org/10.1007/s10529-024-03530-y","url":null,"abstract":"<p>Amides are an important type of synthetic intermediate used in the chemical, agrochemical, pharmaceutical, and nutraceutical industries. The traditional chemical process of converting nitriles into the corresponding amides is feasible but is restricted because of the harsh conditions required. In recent decades, nitrile hydratase (NHase, EC 4.2.1.84) has attracted considerable attention because of its application in nitrile transformation as a prominent biocatalyst. In this review, we provide a comprehensive survey of recent advances in NHase research in terms of natural distribution, enzyme screening, and molecular modification on the basis of its characteristics and catalytic mechanism. Additionally, industrial applications and recent significant biotechnology advances in NHase bioengineering and immobilization techniques are systematically summarized. Moreover, the current challenges and future perspectives for its further development in industrial applications for green chemistry were also discussed. This study contributes to the current state-of-the-art, providing important technical information for new NHase applications in manufacturing industries.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"25 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186271","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Immobilization coupling with aptamer assisted dual cycle amplification for sensitive sEVs isolation and analysis","authors":"Yu He, Ying Ren, Jiawen Tang","doi":"10.1007/s10529-024-03526-8","DOIUrl":"https://doi.org/10.1007/s10529-024-03526-8","url":null,"abstract":"<p>Precise identification of small extracellular vesicles (sEVs) is crucial for improving disease diagnosis and treatments, such as bladder cancer. However, accurate isolation and simultaneously quantification of sEVs remain a huge challenge. We have introduced a new technique that combines immobilization with aptamer-assisted dual cycle amplification to isolate and analyze sEVs with high sensitivity. In this method, the CD9 protein antibody is attached to the plate’s surface for the initial identification of sEVs, while an aptamer probe is used to detect the exosomal surface protein CD63. We have created an sEVs-surface method that combines target recognition initiated signal recycling and rolling circle amplification (RCA) for signal amplification. This approach allows for the “AND” logic analysis of dual biomarkers, enabling both sEVs quantification and tracing. The proposed approach has a broad detection range and a low limit of detection. Moreover, the established method showed good stability in detecting sEVs with a low coefficient of variation. Our method can effectively isolate certain sEVs and accurately identify them, making it suitable for many uses in biological science, biomedical engineering, and personalized medicine.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"5 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Martina Pistek, Peter Andorfer, Reingard Grabherr, Barbara Kraus, Juan A. Hernandez Bort
{"title":"Factors affecting rAAV titers during triple-plasmid transient transfection in HEK-293 cells","authors":"Martina Pistek, Peter Andorfer, Reingard Grabherr, Barbara Kraus, Juan A. Hernandez Bort","doi":"10.1007/s10529-024-03520-0","DOIUrl":"https://doi.org/10.1007/s10529-024-03520-0","url":null,"abstract":"<p>The efficiency of triple-plasmid transfection in recombinant Adeno-Associated Virus (rAAV) production was analyzed by examining two distinct HEK-293 cells lines. These were categorized as high producer (HP) and low producer (LP) based on their differing levels of productivity under identical conditions. Analysis of RNA expression levels of viral genes revealed disparities in plasmid derived gene expression between the cell lines. Further assessment of transfection efficiency utilizing labeled plasmids revealed lower plasmid uptake and less efficient nuclear transport in LP cell line. Additionally, we observed inferior translation activity in LP, contributing to its shortcomings in overall productivity. In our attempt to optimize plasmid ratios to enhance fully packaged rAAV particle yield, we discovered cell-line-specific optimization potential. The findings highlight the transfection's complexity, urging tailored strategies for improved rAAV production based on each cell line's characteristics, enhancing understanding and guiding further efficiency optimization in rAAV production.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"3 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mariana Erthal Rocha, Norberto Mangiavacchi, Marcia Marques, Lia Teixeira
{"title":"Succession from acetoclastic to hydrogenotrophic microbial community during sewage sludge anaerobic digestion for bioenergy production","authors":"Mariana Erthal Rocha, Norberto Mangiavacchi, Marcia Marques, Lia Teixeira","doi":"10.1007/s10529-024-03528-6","DOIUrl":"https://doi.org/10.1007/s10529-024-03528-6","url":null,"abstract":"<p>To assess microbial dynamics during anaerobic digestion (AD) of sewage sludge (SWS) from a municipal Wastewater Treatment Plant (WWTP), a Biochemical Methane Potential (BMP) assay at 37 °C under mono-digestion conditions was conducted. Utilizing the Illumina MiSeq platform, 16S ribosomal RNA (rRNA) gene sequencing unveiled a core bacterial community in the solid material, showcasing notable variations in profiles. The research investigates changes in microbial communities and metabolic pathways to understand their impact on the efficiency of the digestion process. Prior to AD, the relative abundance in SWS was as follows: <i>Proteobacteria</i> > <i>Bacteroidota</i> > <i>Actinobacteriota</i>. Post-AD, the relative abundance shifted to <i>Firmicutes</i> > <i>Synergistota</i> > <i>Proteobacteria</i>, with <i>Sporanaerobacter</i> and <i>Clostridium</i> emerging as dominant genera. Notably, the methanogenic community underwent a metabolic pathway shift from acetoclastic to hydrogenotrophic in the lab-scale reactors. At the genus level, <i>Methanosaeta</i>, <i>Methanolinea</i>, and <i>Methanofastidiosum</i> predominated initially, while post-AD, <i>Methanobacterium</i>, <i>Methanosaeta</i>, and <i>Methanospirillum</i> took precedence. This metabolic transition may be linked to the increased abundance of <i>Firmicutes</i>, particularly <i>Clostridia</i>, which harbor acetate-oxidizing bacteria facilitating the conversion of acetate to hydrogen.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"60 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142224326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Research on the application of heterotrophic nitrification-aerobic denitrification bacteria in membrane bioreactor (MBR)","authors":"Tianrui Zhai, Tiantao Zhao, Yuhao Zhong, Peipei Chen, Guojian Li, Liang Teng, Lijie Zhang, Hao Liu","doi":"10.1007/s10529-024-03529-5","DOIUrl":"https://doi.org/10.1007/s10529-024-03529-5","url":null,"abstract":"<p>Inoculating heterotrophic nitrification-aerobic denitrification bacteria (HN-AD) to enhance membrane bioreactor (MBR) efficiency may result in the loss of functional bacteria. Therefore, this study compares the application results of enhancing MBR with a self-designed biological amplifier coupled with HN-AD against the performance of conventional MBR. After enhancement, the MBR achieved a removal efficiency of 96.7% for NH<sub>4</sub><sup>+</sup>-N (100 mg/L) and 96.4% for COD (400 mg/L) in synthetic wastewater. There was a 33% increase in TN (100 mg/L) removal efficiency. The dominant bacteria in the MBR were <i>Alcaligenes</i> (48.4%) and <i>Thauera</i> (15.2%). Additionally, the abundance of denitrification genes (<i>nir</i>K, <i>nor</i>B, <i>nos</i>Z) increased in the enhanced MBR, contributing to improved TN removal efficiency. The use of a biological amplifier effectively solved the problem of HN-AD loss in sewage treatment.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"9 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142186273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Biotechnology LettersPub Date : 2024-08-01Epub Date: 2024-06-06DOI: 10.1007/s10529-024-03500-4
Tomasz Boruta, Grzegorz Englart, Martyna Foryś, Weronika Pawlikowska
{"title":"The repertoire and levels of secondary metabolites in microbial cocultures depend on the inoculation ratio: a case study involving Aspergillus terreus and Streptomyces rimosus.","authors":"Tomasz Boruta, Grzegorz Englart, Martyna Foryś, Weronika Pawlikowska","doi":"10.1007/s10529-024-03500-4","DOIUrl":"10.1007/s10529-024-03500-4","url":null,"abstract":"<p><strong>Objective: </strong>The aim of this study was to determine the influence of the inoculation volume ratio on the production of secondary metabolites in submerged cocultures of Aspergillus terreus and Streptomyces rimosus.</p><p><strong>Results: </strong>The shake flask cocultures were initiated by using 23 inoculum variants that included different volumes of A. terreus and S. rimosus precultures. In addition, the axenic controls were propagated in parallel with the cocultures. UPLC‒MS analysis revealed the presence of 15 secondary metabolites, 12 of which were found both in the \"A. terreus vs. S. rimosus\" cocultures and axenic cultures of either A. terreus or S. rimosus. The production of the remaining 3 molecules was recorded solely in the cocultures. The repertoire and quantity of secondary metabolites were evidently dependent on the inoculation ratio. It was also noted that detecting filamentous structures resembling typical morphological forms of a given species was insufficient to predict the presence of a given metabolite.</p><p><strong>Conclusions: </strong>The modification of the inoculation ratio is an effective strategy for awakening and enhancing the production of secondary metabolites that are not biosynthesized under axenic conditions.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"601-614"},"PeriodicalIF":2.0,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11217084/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141282895","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}